New mass spectrometry (MS) technologies present an exceptional opportunity for creative applications with potential for breakthrough discoveries. We request funding a high performance Q-TOF mass spectrometer (AB SCIEX TripleTOF" 5600+), interfaced with an ultra-high performance liquid chromatography system (chip-based ekspert" nanoLC 425). This state-of-the-art system will be located in the Proteomics and Mass Spectrometry Facility of the Cornell University Biotechnology Resource Center (BRC). Implementation of the instrument in the BRC as a university-wide shared research resource will enable cost effective access to this vital technology for a broad range of NIH-funded research. There is a growing demand for proteomics and mass spectrometry services at Cornell, as shown by the rapid and continuing growth in the number of facility users and samples processed by the facility over the past nine years. The proposed integrated instrument system will expand the capacity and capabilities of the facility but not duplicate the existing instrumentation at the core facility, which is currently utilized at near full capacity. The need fr increased performance and capacity is particularly critical with respect to applications involving analysis of complex samples which require enhanced scan speed, high sensitivity and resolution, and highly accurate mass measurements. The integrated system requested will provide improved dynamic range and will enable confident identification of peptides from low abundance proteins and of post-translational modifications (PTMs) in complex samples. The TripleTOF 5600+ will significantly enhance the facility's capability to support proteomics discovery studies in qualitative exploration as well as high throughput profiling and high-resolution quantification studies involving complex biological samples. The ekspert" nanoLC 425 with the cHiPLC(R) system is an integrated chip-based UHPLC that provides broad 2D LC flexibility and continuous direct nano flow. It has flexibility in workflow selection using chip-baed columns and offers a broad range of flow rates for both global discovery and higher throughput targeted quantitation. The nanoLC 425 system will provide automated sample handing and high resolution peptide separation prior to MS, which will enable continuous 24/7 analysis of very complex samples. The BRC has extensive experience in running LC MS instrument systems and providing MS applications as core facility services. The proposed instrument system will be readily integrated into the facility's established proteomics workflows and will significantly expand the facility's quantitative proteomics pipeline. Placement of the instrument system in the BRC will enable a diverse group of NIH funded investigators to undertake research projects that would not otherwise be economically or technically feasible. This proposal describes 39 projects from 16 NIH funded investigator groups that will utilize the proposed system. The requested instrument will benefit a wide range of current and future NIH-funded projects.
The instrument system requested has technical capabilities that exceed those possessed by the existing instruments in the Cornell BRC that are currently available to 16 investigators with 39 NIH funded projects, and would facilitate accomplishing the goals of these projects by enabling significant improvements in both the quantity/quality of data while reducing turnaround times. The proposed instrumentation will play a crucial role in advancing knowledge and understanding in a wide variety of health related research, including studies of cancer, Parkinson's disease, insulin resistant syndromes, infertility, cancer transformation, Kallman's Syndrome, toxicity mechanism of dioxin, and in the development of glycotherapeutic applications.
|Zhang, Xiaoyu; Khan, Saba; Jiang, Hong et al. (2016) Identifying the functional contribution of the defatty-acylase activity of SIRT6. Nat Chem Biol 12:614-20|
|Martin, Laetitia B B; Sherwood, Robert W; Nicklay, Joshua J et al. (2016) Application of wide selected-ion monitoring data-independent acquisition to identify tomato fruit proteins regulated by the CUTIN DEFICIENT2 transcription factor. Proteomics 16:2081-94|
|Wang, Jie; Sevier, Carolyn S (2016) Formation and Reversibility of BiP Protein Cysteine Oxidation Facilitate Cell Survival during and post Oxidative Stress. J Biol Chem 291:7541-57|